Myophysics basic knowledge and detection methods: myosome culture technology!

basic knowledge and detection methods: myophysics,
training
technology!
mycomal
also known as mold, is a very small primary nuclear cell, completely without cell walls. The shape of the bacterium is variable, with a high degree of polygonality, spherical, pear-shaped, branched or spiral silky, etc. Nutritional requirements are demanding, growing on complex
cultures
with high
serum
, and the bacteria on solid cultures have a characteristic appearance of fried eggs.
culture base: bovine heart soup compound culture base, etc.
isolated culture: sterile to take lung, liver, spleen, kidney tissue, cut directly into small pieces placed on a solid culture base to draw a straight line, and then with sterile platinum earrings in its vertical direction, or the tissue block cut into a smooth oar, after continuous dilution of the liquid culture base culture. If the liquid material is cow's milk, excreta, fetal fluid, etc., you can directly take
0.2ML inoculated in liquid culture or 0.1ML inoculated in solid culture. Solid
agar
flat set 37 degrees Celsius humid oxygen demand conditions or culture 48-96h under 5% carbon dioxide, with oblique light or magnify 25-40 times stereo
scope
check, if there are specific bacteria appear, should choose scattered in a single bacteriospheric knife cut small agar block, with a pasteurized straw carefully Drop and surround agar inhalation, and then move it into a liquid broth culture tube containing 2-3ML culture 48h, its culture through 0.45 micro-liter aperture size membrane
filtration
after 1:10 and 1:100 dilution, each dilution of 0.05ML coated on several agar plates, you can obtain pure fuse culture.
compound culture base:
bovine heart soup
1000ml
protein
10g
sodium chloride
5ml
Yeast extract
10ml
sterile horse serum
200ml
penicillin
200IU/ml
experiment
MP
pneumonia mycosm
culture experimental method:
1, specimen collection
progenitors often invade the mucous membrane surface of humans and animals, generally can be applied with cotton swabs after sampling into
2 to 3 ml of mycobacterial liquid medium in a small tube, or take its secretion, if the specimen is tissue block, can be sterilized after the milk or glass homogenization, should be inoculated as soon as possible.
2, separation culture
1, mycosm solid media inoculation method rotation swab to squeeze out the specimen liquid as far as possible, with sterile dropper suction 1 to 2 drops inoculated on the flat dish covered, with L stick smear or tilt rotation flat dish so that the specimen is evenly distributed, placed 37 degrees C incubation.
2, myophysics semi-solid medium inoculation method mixed method: in the preparation of semi-solid medium, medium
heating
dissolved, the temperature dropped to 50 degrees C, add animal serum, yeast immersion After mixing with penicillin, to be cooled to 37 to 40 degrees C, with sterile droppers to absorb specimens 0.5 to 1 ml added to the medium, fully mixed, solidified after 37 degrees C incubation.
3, puncture vaccination method: method with bacterial vaccination method.
, the results of observationThe vast majority of myogens are anaerobic, and a few grow well in atmospheric environments containing
10% CO2 and relative humidity of 80 to 90%. Myloid growth is slow, most in 3 to 4d growth of bacteria, a few in 1 to 2w square growth of typical bacterios, in flat dishes solid media growth on the bacteria in the shape of "oil omelettes", the edge is not neat.
Ivy, Appendix
Sugar Enzyme Culture (pneumonia myophylitis media)
Basic media
70ml
horse (or bovine serum)
20ml
25% yeast immersion 10ml
0.4% phenolic red 0.5ml
penicillin
10000IU/ml 5.0ml
tune
pH 7.8
pneumonia mycopheric culture is very simple, especially liquid culture, do not need protection!
the recipe is a bit complicated and vague, simple:
PPLO 21g (commoditized buy)
yeast powder
6g
horse serum
200ml (no use calf serum instead)
L-cysteine 0.3g

glucose
10g
1% phenol red 4ml
AMP several (0.1-0.2g, plus dots without effect)
super sometimes plus
SMZ and TMP!
ph to tune, about 7.5-7.6!
red can be high pressure!
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